Download Izabella Battonyai

UNIVERSITY OF PÉCS
Biological Doctoral School
Ultrastructural, functional and molecular
aspects of the organization of the olfactory
center of gastropod species (Helix pomatia,
Limax valentianus)
PhD Thesis
Izabella Battonyai
Supervisor:
Károly Elekes PhD
Doctor of Biological Sciences
PÉCS, 2014
Ultrastructural, functional and molecular
aspects of the organization of the olfactory
center of gastropod species (Helix pomatia,
Limax valentianus)
PhD Thesis
Izabella Battonyai
Supervisor:
Károly Elekes PhD
Doctor of Biological Sciences
PÉCS, 2014
1. INTRODUCTION
In terrestrial pulmonata snails olfaction is the main sensory modality,
it determinates and influences many specific behavior (Stocker 1993). We
have immersed knowledge about their olfactory system, including its
structural and functional organization (Gelperin, 1999, Chase, 2002). Both
electrophysiological and autoradiographical methods supported that the
receptor cells, located on the tip of the tentacles take part not only in
mechanosensory but in olfactory information processing (Chase 1982). The
olfactory epithelium is situated in the ventral part on the tip of the tentacles,
from where through the receptor cells the odor information is is sent to and
integrated in the tentacular ganglion followed by the information processing
processes in the CNS. The center of odor processing is the procerebrum
(olfactory lobe, PC), which is located in the cerebral ganglion and is a
characteristic unit in Stylommatophora (Pulmonata) snails (Van Mol 1974).
There are also a number of morphological and physiological data proving
that the PC takes part in olfaction processes, it has important role in
olfactory information processing and in odor learning and memory formation
as well (Gelperin, 1999; Chase, 2002). Although in early electron
microscopic studies Zs.-Nagy and Sakharov (1969) identified synapses in
the cell layer of the PC, our knowledge is incomplete regarding the exact
ultrastructure and synaptology of the cell body layer.
Apart from some neuropeptides and the octopamin, neurotransmitters
in the Gastropoda nervous system are similar to that can be found in the
vertebrates (e.g. acetylcholine, serotonin, dopamine, glutamate) (Chase
2002). Among them, serotonin (5-HT) has been known for a very long time
as an important signal molecule, which is widely distributed in invertebrates
(Gerschenfeld, 1973; Gardner and Walker, 1982; Walker et al., 1996).
In the Gastropod nervous CNS it has been revealed that many 5HTergic processes are present and arborizing in the medullary neuropil of
the PC (Osborne and Cottrell, 1971; Inoue et al., 2004; Matsuo et al., 2009).
However, the high-resolution analysis of the organization and intercellular
interacions of the 5-HT containing elements within the PC, including the
identification of their postsynaptic targets, is still missing.
Potassium (K+) channels can be found virtually in all living organism,
responsible for maintaining the electronic activity of the cells (Kurachi et al.
1999). Voltage-dependent potassium channels are the largest ion channel
family. They play a pivotal role in bursting of nerve cells, neuronal
excitability and re-polarizing cells following action potentials (Hille, 2001;
Conley and Brammar, 1999). The first K+-current was recorded from
neurons of the slug Onchidium by Hagiwara et al. (1961). Although, by
immunohistochemistry (IHC), Azanza et al. (2008) demonstrated the
presence of different ion channels in the neurons of the subesophageal
ganglion complex of Helix aspersa, the detailed investigation of K+-channels
in the entire CNS, regarding their precise distribution, projection patterns as
well as the visualization of possible intercellular contacts established by K+channel containing neurons have completely been neglected. The precise
localization and functional characterization of K+-channels would be
indispensable to have a better insight in their possible implication in signal
transduction processes in the nervous system of gastropods.
2. AIMS
It is clear that still many information and details are missing
concerning the olfactory lobe (procerebrum, PC) of the terrestrial pulmonate
snails, including its anatomical, ultrastructural organization and its synaptic
connections. Moreover, the exact organization of the 5-HTergic innervation
pattern and its synaptology is not known.. Data and the possible functional
interpretation have also been missing on the presence and cellular
localization of K+-channels in the gastropod CNS. In the globuli cells of the
olfactory center, PC, the precise localization, functional characterization and
synaptic connections of K+-channels seem to be indispensable to have a
better insight in voltage-gated K+-channels as to their involvement in signal
transduction processes in the nervous system of gastropods, including odor
information processing.
The aims of our present work were the following.
1. To revise and define the ultrastructural organization of the PC cell body;
layer, including the local neuropil areas and the intercellular connections
2. To describe the 5-HT-IR innervation system of the PC in details, with
special attention to the cell body layer and the different forms of intercellular
connections;
3. To identify voltage-gated K+-channels in the CNS, including the PC, and
possibly reveal their functional characteristics.
3. MATERIALS AND METHODS
3.1. Animals
Adult specimens of the garden (land) snail Helix pomatia and the slug
Limax valentianus were used. Specimens of Helix pomatia were collected in
the surrounding areas, kept thereafter under laboratory conditions and fed on
lettuce. Slugs 11–14 weeks after hatching were maintained under laboratory
conditions and fed on a mixed diet of rat chow (Oriental Yeast, Tokyo,
Japan), wheat starch (Wako, Osaka, Japan) and vitamins (Oriental Yeast).
3.2. Electron microscopy
For fixation, the CNS of both species were isolated and pinned out in
a Petri-dish and covered with a mixture of either 2.5% PFA and 1 % GA.
Preparations were post-fixed in 1% OsO4, dehydrated in graded ethanol and
propylene oxide, and finally embedded in Araldite. Block staining was
performed in 70% ethanol saturated with uranyl acetate. For orientation 1
µm semi-thin sections were stained with toluidine blue. 50-60 nanometer
ultrathin serial sections from the PC region were taken with an LKB
Novacut ultramicrotome, stained with lead citrate, and viewed in a JEOL
1200 EX, a JEOL 1200 EXII, and a Hitachi H-7650 electron microscope.
3.3. Immunohistochemistry
In our experiments we used two-step indirect (fluorescent dye or
peroxidase-IgG) or three-step avidin-biotin complex (ABC) for
visualization. In case of 5-HT IHC only the cerebral ganglia (PC), while in
case of the ion channels the whole CNS were fixed in 4% PFA and cut into
16 µm slices on a cryostat. The following primary antibodies were used:
monoclonal anti-5-HT (1:500-1000); polyclonal anti-Kv3.4 and anti-Kv4.3
(1:500 vagy 1:1000); and monoclonal mouse anti-Kv2.1 (1:500). After this
the next secondary antibodies were used depending the primary antibody:
rabbit anti-donkey IgG conjugated with FITC or TRITC (1:200), or biotinconjugated goat anti-mouse igG or anti-rabbit IgG (1:200), following avidinHRP (1:200) incubation. The HRP-reaction was visualized by using 0.05 %
3.3’-DAB and 0.01% H2O2. The sections were mounted in 1:1 mixture of
glicerin:PBS and viewed in a Zeiss Axioplan compound microscope attached
to a Canon PS G5 digital camera or CCD (Alpha DCM510, Hangzhou
Scopetek Opto-Electric) camera. The specificity of each antibody was tested
by applying both negative (omitting the primary antibody) and preabsorption control experiments. No immunostaining could be observed
following these control experiments.
3.4. Correlative light- and electron microscopic immunocytochemistry
Helix and Limax CNS were dissected and fixed with 2.5 % PFA and
0.1 % GA, than the PC –s (5-HT, Kv4.3) and the caudo-medal lobe of the
pedal ganglia (Kv4.3) were postfixed in 10% PFA and cut into 50 µm slices
with a Vibratome. Slices were incubated in mouse monoclonal anti-5-HT
antiserum or rabbit polyclonal anti-Kv4.3 antiserum. In case of anti-5-HT
the slices were processed for a two-step peroxidase immunocytochemistry
(HRP-DAB), the secondary antibody concentration was 1:50. In case of
Kv4.3 the immunoreaction was visualized with a one-step polimer-HRP IHC
detection system. The slices were post-fixed in 1% OsO4, dehydrated, and
then mounted on slides in Araldite. After polymerization, the slices were
analyzed in a light microscope. Following light microscopy, slices
displaying high quality immunolabeling were selected and re-embedded for
electron microscopy and Camera lucida reconstruction also have been made.
Ultrathin sections of 50–60 nm were cut, stained with lead citrate, and
viewed in the electron microscopes.
3.5. Western blot experiments
For Kv2.1 and Kv3.4 immunoblotting 10 pieces of the CNS including
the surrounding neural sheath or 10 pieces of PC, meanwhile for Kv4.3
immunoblotting 10 pieces of desheathed CNS, separated connective tissue
sheath and PC, respectively, were homogenized in an SDS containing
lysating buffer. Electrophoresed proteins were blotted onto nitrocellulose
membranes and membrane strips were incubated with the primer antisera
(anti-Kv2.1, anti-Kv3.4 and anti-Kv4.3, 24h) than HRP-conjugated to
seconder antibodies. The visualization of the reaction product was carried
out with an enhanced chemilumescent (ECL) reagent, or in Tris–HCl buffer
(pH 7.6) containing 0.05% DAB (Sigma) and 0.01% H2O2. Method control
experiments were performed by omitting the primary antibody and
preabsorption specificity control was made with the recognizing peptides.
3.6. Electrophysiology
3.6.1. Patch-clamp experiments
Perforated patch-clamp recordings on Limax PC globuli cells were
made using previously published experimental protocols (Watanabe et al.
2003). Focal application of 5-HT was performed by a glass micropipette
placed beside the soma (≤50 μm). 5-HT was dissolved in Limax saline
solution containing 0.05 %, Fast Green (Sigma) to allow visual confirmation.
In Helix PC, electrophysiological experiments were made on neurons of the
Helix PC selected from those expressing immunolabeling against Kv4.3 and
Kv2.1. Current recording was performed under an inverted microscope
(Leitz Labovert FS, Germany) using tight seal whole cell patch-clamp
recording. For data acquisition and clamp protocols, the amplifiers were
connected via a TL DMA interface to a computer supplied with pClamp 5.5
software. Currents were separated using special voltage protocols.
3.6.2. Voltage-clamp experiments
Further electrophysiological experiments were made on neurons of the
Helix CNS expressing immunolabeling against Kv4.3 (pedal ganglion,
caudomedial lobe neurons), Kv2.1 (PC), and Kv3.4 (buccal ganglion, B2 cell)
channels, respectively. Current recordings were performed using a
GeneClamp amplifier in two microelectrode voltage-clamp mode. Data
acquisition and analysis were performed using Digidata interface and
pCLAMP software (Axon Instruments). In part of the experiments for
recording transient outward K-current the physiological solution was
modified, containing 5 mM tetraethylammonium chloride (TEA, Sigma,
Budapest) and NaCl was replaced for sucrose. In sucrose and 5 mM TEA
saline, the Na-dependent inward current and part of the K-currents, except
A-current, were eliminated. For blocking the following drugs were used:
blood-depressing substance (BDS-II, Alomone Labs), 4-aminopyridine (4AP) and 20 mM TEA (both from Sigma).
4. RESULTS
4.1. Ultrastructure and synaptology of the cell body layer of the
procerebrum
In semi-thin section taken from both the Helix and Limax PC local
neuropils were present in the entire depth the globuli cell body layer
globulus. At ultrastructural level, these neuropil regions were of different
size and they display a gradual branching, in the course of which the smaller
neuropil units contained only a few axon profiles, meanwhile forming a
more intimate contact with the surrounding globuli cell bodies. Finally,
individual axons established axo-somatic contacts.
At the ultrastructural level, the form of the axo-somatic contacts in
the PC cell body layer widely varied. Both specialized and unspecialized
axo-somatic contacts were established by varicosities deeply embedded into
the globuli cells perikarya. The axo-somatic and axo-axonic contacts
observed in the local neuropils of both the Helix and Limax PC revealed
different forms of synaptic configurations, such as synaptic divergence and
presynaptic modulation. In addition, along the unspecialized close contacts
exocytotic membrane configurations could also be observed, referring to the
active release of signal molecules.
4.2. The chemical-neuroanatomy of the 5-HTergic innervation in the
procerebrum
In 50 μm Vibratome slices taken from the Helix PC the characteristics
of the 5-HT-immunoreactive (IR) innervation pattern has been visualized,
establishing the determining role in the innervation of a thick labeled axon
bundle of extrinsic (metacerebral) origin. Entering the PC, the 5-HT-IR
fibers showed a rich arborization innervating the entire olfactory lobe. This
pattern from broad modulatory system might be involved both in processing
the olfactory information and execution of proper behavioral responses given
to odor stimuli. The individual axo-somatic innervation of the gloubli cells
refers to the local role of 5-HT.
By camera lucida tracing the until unknown 5-HT-IR innervation of
the cell body layer of the Limax PC-ben could also be visualized, and in
addition the functional presence of 5-HT has also been proved in this system
by demonstrating the 5-HT sensitivity of the spiking (B) cells, suing patchclamp mode, following the local application of 1 mM 5-HT.
The ultrastructural characteristics of the 5-HT-IR elements innervation
the Helix olfactory have been revealed by correlative light- and
electronmicroscopic immunohistochemical investigations. The 5-HT-IR
varicosities were shown to form frequently close but unspecialized
membrane contacts with perikarya and agranular axon profiles. They formed
with them synaptic configurations as well.
4.3. Immunohistochemical identification and distribution of voltage
gated K+-channels in the Helix CNS
4.3.1. Distribution of Kv2.1 immunoreactive elements
Kv2.1-IR neurons could be observed in all ganglia of the CNS. In the
PC numerous Kv2.1-IR globuli cells were present. At higher magnification
small immunopositive varicosities were seen, surrounding unlabeled globuli
cells. In the labeled cells the immunoreactivity was frequently concentrated
only in a smaller part of the cytoplasm. The majority of the Kv2.1-IR
neurons projected to the medullary neuropil.
4.3.2. Distribution of Kv3.4 immunoreactive elements
Kv3.4-IR neurons have been found in the entire Helix CNS. The
cytoplasm of the labelled cells contained dot-like precipitates that were
frequently localized at the periphery, nearby the cell membrane. The axon
processes of the labeled neurons projected to the neuropils, where they
arborized. Kv3.4 immunoreactivity was displayed by a single giant neuron,
the B2 cell in the buccal ganglion. In the PC, a few Kv3.4-IRglobuli cells
could be identified, located along the borderline between the cell body layer
and the terminal medulla.
4.3.3. Distribution Kv4.3 immunoreactive elements
Kv4.3 immunoreactive neurons occurred in the entire CNS but the
visceral ganglion, sending several labeled fibers to the neuropils. K v4.3-IR
elements have also been identified in non-neuronal tissue as well,
concentrated in the connective tissue sheath, surrounding the ganglia, and in
the aorta wall. In the buccal ganglion, the B2 motoneuron was labeled again,
in addition to a significant group of neurons of larger size. In the PC, the
immunopositive globuli cells were partly distributed throughout the cell
body layer, and partly lined up along the borderline between the cell body
layer and the medullary neuropil. In the cell body layer, labeled axon
processes were collected in septum-like bundles running among the cell
bodies. The subesophageal ganglion complex contained numerous Kv4.3-IR
cell groups, mainly located in the pedal and pleural ganglia. At
ultrastructural level, the labeled elements were the same type both in the PC
and the pedal ganglion, containing numerous 80-100 nm granular vesicles.
The character of the labeling was also similar, showing dot-like precipitates
on the surface of the granular vesicles and/or distributed in the axoplasm.
4.4. Identification of the ion channels by Western blot
In the case of the Kv2.1, Kv3.4 channels the PC and the whole CNS,
whereas in the case of the Kv4.3 channel the homogenates of the ganglionic
connective tissue were also analyzed, regarding the presence of the ion
channels. After blotting the specific bands appeared at the molecular weights
expected on the basis of the literature data. In case of Kv2.1 antibody it was
110 kDa, for Kv3.4 antibody it was found at 240 kDa, and the Kv4.3 antibody
showed labeling at 73 kDa.
4.5. Electrophysiological characterization of K+-channels in the central
neurons of Helix
In the B2 neuron of the buccal ganglion displaying Kv3.4
immunoreactivity a fast, A-type K+-current was registered. The current could
be inactivated by adding 5 mM TEA, that blocks the inward Na+- and the
delayed rectifier Ca2+-activated K+-currents. The A-type current could also
be blocked effectively with 4-AP and BDS-II, whereas delayed rectifier
currents were blocked by 20 mM TEA However, low concentration (4 mM)
4-AP did not result in significant changes. From the neurons in the caudomedial lobe of the pedal ganglion characterized by K v4.3 immunreactivity
another type of transient A-type K+-current could be recorded, having a
smaller amplitude and a slower kinetic. On the other hand, its voltagecurrent character resembled very much that of the fast transient A-current.
This current type could also be blocked by 4-AP, whereas it was less
sensitive against TEA. K+-currents were successfully recorded from the PC
globuli cells both in voltage-clamp, and patch-clamp mode. Using voltage-
clamp technique, the K+-currents of globuli cells were analyzed that
displayed Kv2.1 and Kv4.3 immunoreactivity, respectively. In a part of the
slow, A-type K+-current was recorded, although a delayed rectifier current
component could also be observed.
5. SUMMARY
The procerebrum (PC) of terrestrial pulmonates is the center of odor
processing. We have little information about the ultrastructure organization
of the cell body layer and the 5-HTerg innervation system of the PC at high
resolution. Therefore, we have performed both routine electron microscopy
and correlative light- and electron microscopic immunohistochemistry, in
order to resolve the fine structural organization, and 5-HT immunoreactive
innervation of the PC. Data have also been sporadic on the presence,
distribution and cellular localization of ion channels, especially the K +channels in the snail CNS. To have a better insight in the distribution of K +channels, we have applied immunohistochemical and electrophysiological
methods. The precise localization and functional characterization of K+channels would be indispensable for our understanding of cell-to-cell
signaling processes in the nervous system of gastropods, important models
of comparative neurobiology.
Our results can be summarized as follows:
1. We have described the detailed ultrastructural organization in the globuli
cell layer of the PC in two prominent terrestrial pulmonate gastropod
species, Helix pomatia and Limax maximus. We identified local neuropil
regions of different size which are characterized by axo-axonic and axosomatic contacts, displaying different forms of synaptic configurations,
(divergence and presynaptic modulation). Along unspecialized axo-somatic
contacts exocytotic membrane configurations were also observed, indicating
the process of extrasynaptic release. These observations widen the
interpretation possibilities concerning the modulation of integrative olfactory
processing networks in the PC.
2. Immunohistochemistry revealed a rich innervation established by a 5-HTIR network in the Helix PC. The cell body layer received a massive
innervation by varicose axon processes, forming a perisomatic basket-like
arrangement. The neuropil regions also contained many 5-HTergic
processes. At ultrastructural level, 5-HT-IR fibers often formed axo-somatic
and axo-axonic connections in the PC, establishing different synaptic
configurations. Our results suggest that 5-HT is a pivotal signal molecule in
the PC, having a general modulatory role.
3. Immunohistochemical experiments revealed neurons displaying labeling
against three voltage-gated potassium channels (Kv2.1, Kv3.4 and Kv4.3)
throughout the Helix CNS. The distribution pattern of the different channel
expressing cells was similar. The antibodies used labeled distinct sets of
neurons, which innervated the neuropil of the ganglia. The overall discrete
number of labeled cell groups suggests a well-defined specific role for each
K+-channels studied. At ultrastructural level we have identified Kv4.3-IR
varicosities in some part of the Helix CNS (PC, pedal ganglion). The labeled
varicosities were characterized by a rather uniform fine structure, which also
suggests a different role of K+-channels in modulating processes.
4. Patch-clamp and voltage-clamp experiments have proven the functional
existence of the different K+-channels in the Helix CNS. We successfully
identified a typical fast transient K+-current (A-current) in the Kv3.4-IR B2
neuron of the buccal ganglion. Using the same voltage-protocol and the
same saline, different transient (slower, A-type) K+-current was recorded
from neurons located in the caudo-medial lobe of the pedal ganglion. In the
PC, delayed rectifier K+-current in neurons expressing Kv2.1 channel, and
transient A-type K+-current in neurons expressing Kv4.3 channel have been
demonstrated. It is assumed that both ion channels are involved in odor
information processing.
6. List of publications and presentations
6.1. Publications
Battonyai I, Krajcs N, Serfőző Z, Kiss T, Elekes K. (2014) Potassium
channels in the central nervous system of the snail, Helix pomatia:
localization and functional characterization Neuroscience 268:87-101 IF:
3,389
Battonyai I, Serfőző Z, Elekes K. (2012) Potassium channels in the Helix
central nervous system: preliminary immunhistochemical studies. Acta Biol.
Hung. 63 (Suppl. 2), 146-150 IF: 0,504
Battonyai I, Elekes K. (2012) The 5-HT immunoreactive innervation of the
Helix procerebrum. Acta Biol. Hung. 63 (Suppl. 2), 96-103. IF: 0,504
Elekes K, Battonyai I, Kobayashi S, Ito E. (2013) Organization of the
procerebrum in terrestrial pulmonates (Helix, Limax) reconsidered: cell mass
layer synaptology and its serotonergic input system, Brain Struct. Funct.
218:477-490 IF: 7.837
Pirger Z, Battonyai I, Krajcs N, Elekes K, Kiss T (2013) Voltage-gated
membrane currents in neurons involved in odor information processing in
snail procerebrum. Brain Struct. Funct. Brain Struct. Funct. 219:673–682
IF: 7.837
6.2. Oral presentations and posters
Battonyai I, Krajcs N, Kiss T, Elekes K.: Potassium channels in the central
nervous system of the snail, Helix pomatia, XIV. MITT Konferencia,
Budapest, január 17-19., 2013
Battonyai I, Elekes K.: Synapses and neurotransmitters in the olfactory
center of the snail, 7th European Conference on Comparative Neurobiology,
Budapest April 25-27, 2013
Battonyai I, Krajcs N, Kiss T, Elekes K: Distribution of voltage-gated
potassium channels in the snail central nervous system, FFRM, Prague,
September 11 - 14, 2013
Battonyai I, Krajcs N, Serfőző Z, Kiss T, Elekes K: Potassium channels in
the central nervous system of the snail, Helix pomatia. MITT 14th
Conference of the Hungarian Neuroscience Society, January 17-19, 2013,
Budapest, Hungary. Abstract: MITT Abstract Book, ISBN 978-963-88224-20, p. 84-85.
Krajcs N, Battonyai I, Hernádi L, Elekes K, Kiss T: Neural control of string
muscles responsible for patterned tentacle movements of the snail, Helix
pomatia. International IBRO Workshop, 2012. január 19-21, Szeged.
Abstract: Ideggyógy. Szemle/Clin. Neurosci. 2012;65(S1):39
Battonyai I, Serfőző Z, Krajcs N, Kiss T, Elekes K: Immunohistochemical
visualization of potassium channels in the central nervous system of the
snail. 8th FENS Forum of Neuroscience, July 14-18, 2012, Barcelona, Spain
Battonyai I, Elekes K: Is serotonin a pivotal modulator in the snail olfactory
center? MITT 13th Conference of the Hungarian Neuroscience Society,
January 20-22, 2011, Budapest, Hungary. Abstract: Front. Neurosci.
Conference Abstract: 13th Conference of the Hungarian Neuroscience
Society (MITT). doi: 10.3389/conf.fnins.2011.84.00094
Battonyai I, Serfőző Z, Elekes K: Potassium channels in the Helix central
nervous system: preliminary immunhistochemical studies. 12th Symposium
on Invertebrate Neurobiology (ISIN), August 31 – September 04 2011,
Tihany, Hungary
Battonyai I, Elekes K: The 5-HT immunoreactive innervation of the Helix
procerebrum. 12th Symposium on Invertebrate Neurobiology (ISIN), August
31 – September 04, 2011, Tihany, Hungary
Battonyai I, Krajcs N, Kiss T, Elekes K: Serotonin is involved in both the
central and peripheral regulation of snail olfaction. 3rd European Synapse
Meeting, October 13-15, 2011, Balatonfüred, Hungary
Elekes K, Fekete ZN, Battonyai I, Mikite K, Kobayashi S, Ito E:
Organization of the olfactory center of terrestrial snails (Helix, Limax)
revisited: massive axo-somatic innervation of the globuli cells in the
procerebrum. International IBRO Workshop 2010, Pécs, 2010. január 21-23.
Abstract: Front. Neurosci. Conference Abstract: IBRO International
Workshop 2010. doi: 10.3389/conf.fnins.2010.10.00034
Pirger Z, Kiss T, Battonyai I, Elekes K: Sodium-channel and membrane
current characteristics of the procerebral neurons of Helix pomatia.
International IBRO Workshop 2010, Pécs, 2010. január 21-23. Abstract:
Front. Neurosci. Conference Abstract: IBRO International Workshop 2010.
doi: 10.3389/conf.fnins.2010.10.00117
6.3. Other publications
Kiss T, Battonyai I, Pirger Z. (2014) Down regulation of sodium channels
in the central nervous system of hibernating snails. Physiol. Behav. 131:9398 IF: 3.160
Talapka P, Bódi N, Battonyai I, Fekete E, Bagyánszki M. (2011)
Subcellular distribution of nitric oxide synthase isoforms in the rat
duodenum. World J Gastroenterol. 17:1026-1029. IF: 2.471
Bódi N, Battonyai I, Talapka P, Fekete E, Bagyánszki M. (2009) Spatial
pattern analysis of nitrergic neurons in the myenteric plexus of the
duodenum of different mammalian species. Acta Biol. Hung. 60:347-358.
IF: 0.593